Functional differences and regulation of gene expression of phosphoribosylpyrophophate synthetase isozymes

磷酸核糖焦磷酸合成酶同工酶的功能差异及基因表达调控

• 批准号: 63570109
• 负责人: TAIRA Masanori
• 金额: $ 1.34万
• 依托单位: Chiba University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1988
• 资助国家: 日本
• 起止时间: 1988 至 1989
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-63570109/
• 关键词: Phosphoribosylpyrophosphate; Phosphoribosylpyrophosphate synthetase; Isozyme; Gene expession; Chromosome mapping; cDNA sequence; Gene structure; Initiation codon; 組織特異的発現; 精巣特異的発現; ヌクレオチド合成; cDNAクローニング; アイソフォーム; 組み換えDNA

1. Cloning of CDNA coding rat phosphoribosylpyrophosphate synthetase revealed two distinct types of subunit, referred to as PRS I and PRS II. Tissue-specific expression of PRS I and PRS II genes was shown. In the testis, an additional transcript was noted.2. The chromosomal localization of human PRS I and PRS II genes was determined. In addition, two PRPS1 (PRS I gene)-related genes (PRPS1L1 and PRPS1L2) were found.3. Complete cDNA sequence of rat PRS I and PRS II were determined.4. Genomic DNA corresponding to rat PRPS1 was cloned and characterized. The gene consisted of seven exons.5. cDNA for testis-specific human phosphoribosylpyrophosphate synthetase was cloned and characterized. The gene was located at chromosome 7. The mRNA had a non-AUG initiation codon, ACG.6. cDNAs for rat PRS I and PRS II were inserted into the vector and expressed in Escherichia coli. The enzyme activities of E. coli cells transformed with the expression vectors were 10-20 fold greater than that of untransformed E. coli cells.

1.克隆编码大鼠磷酸核糖焦磷酸合成酶的cDNA揭示了两种不同类型的亚基，称为PRS I和PRS II。显示了PRS I和PRS II基因的组织特异性表达。在睾丸中，注意到额外的转录本。确定了人PRS I和PRS II基因的染色体定位。此外，还发现了两个PRPS 1（PRSI基因）相关基因（PRPS 1 L1和PRPS 1 L2）.测定了大鼠PRS Ⅰ和PRS Ⅱ的cDNA全序列.对大鼠PRPS 1基因组DNA进行克隆和鉴定。该基因由7个外显子组成。睾丸特异性的人磷酸核糖焦磷酸合成酶的cDNA克隆和表征。该基因位于7号染色体上。mRNA具有非AUG起始密码子ACG.6。将大鼠PRS I和PRS II的cDNA插入载体中并在大肠杆菌中表达。酶活测定表明，E.转化的大肠杆菌细胞比未转化的大肠杆菌细胞的转化率高10-20倍。coli细胞。

项目成果

Taira,M.et al.: "Localization of human phosphoribosylpyrophosphate synthetase subunit I and II genes(PRPS1 and PRPS 2)to different regions of the X chromosome amd assignment of two PRPS1-related genes to autosomes." Somatic Cell Mol.Genet.15. 29-37 (1989)

Taira,M.等人：“将人类磷酸核糖焦磷酸合成酶亚基 I 和 II 基因（PRPS1 和 PRPS 2）定位到 X 染色体的不同区域，并将两个 PRPS1 相关基因分配到常染色体。”

Taira, M. et al.: "Nucleotide and deduced amino acid sequences of two distinct cDNAs for rat phosphoribosylpyrophosphate synthetase." J. Biol. Chem., 262 14867-14870, 1987.

Taira, M. 等人：“大鼠磷酸核糖焦磷酸合成酶的两种不同 cDNA 的核苷酸和推导的氨基酸序列。”

Iizasa,T.et al.: "Molecular cloning and sequencing of human cDNA for phosphoribosylpyrophosphate synthetase subunit II." FEBS Lett.244. 47-50 (1989)

Iizasa,T.et al.：“磷酸核糖焦磷酸合成酶亚基 II 的人类 cDNA 的分子克隆和测序。”

Taira, M. et al.: "Tissue-differential expression of two distinct genes for phosphoribosylpyrophosphate synthetase and existence of the testis-specific transcript." Biochim. Biophys. Acta, 1007 203-208, 1989.

Taira, M. 等人：“磷酸核糖焦磷酸合成酶的两个不同基因的组织差异表达以及睾丸特异性转录本的存在。”

Ishijima,S.,et al.: "Complete cDNA sequence of rat phosphoribosylpyrophosphate synthetase subunit I(PRS I)" Nucleic Acids Res.17. 8860 (1989)

Ishijima,S.,et al.：“大鼠磷酸核糖焦磷酸合成酶亚基 I (PRS I) 的完整 cDNA 序列”核酸研究 17。